I would recommend using MFI data to determine level of receptor expression in your whole cell population, but if you have two distinct cell populations, would recommend using proportions (% of positive cells) in regard to your controls such as unstained cells.

Hello Yasmine, You could try using reference beads. Article Performance of calibration standards for antigen quantitatio...

Have a look Roy Carvers's flow cytometry menu

https://biotech.illinois.edu/flowcytometry/protocols/surface-antigen-quantification.

Hope this helps

martin

Dear Yasmine,

I suggest you to look up this article published in Scientific Reports last year.

Le Lann L et al., Standardization procedure for fow cytometry data harmonization in prospective multicenter studies

https://www.nature.com/articles/s41598-020-68468-3.pdf?origin=ppub

Also, look up ISAC web site for MFI calculation and detailed information.

Best wishes,

Dear Yasmine,

You just need to multiply the percent positive cells and mean/median value of your antigen of interest. I would recommend to use the median value rather than mean value as outliers won't effect the median value.

Best Wishes!

Median values should be used for non-gaussian data.